Epitope mapping of SAK by gene-targeted random fragment library
- VernacularTitle:利用目的基因随机表位库筛选葡激酶抗原决定簇
- Author:
Xiaowen WANG
;
Ziyi YANG
;
Jinfeng WANG
;
Xin CAI
;
Tao XU
;
Minji ZOU
;
Jiaxi WANG
- Publication Type:Journal Article
- Keywords:
Straphylokinase;
Epitope;
Gene-targeted random epitope library
- From:
Chinese Journal of Immunology
2000;0(11):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To map dominant antigenic determinants on SAK by gene-targeted fragmemt library . Methods:①The PcAbs specificly against SAK were produced by immunning BALB/C mice. Pu rified the antisera through a SAK-Sepharose 4B affinity chromatography column. The purified PcAbs were biotinylated for next step. ② After constructing SAK r andom epitope library we sequenced 12 isolated clones randomly to ensure its int egrity ,capability and randomness.③The library was screening by situ-clone hy bridization.④Constructed mSAK ,the A1 region deleted mutant of SAK ,and used Western-blot assay to identified its immunoreactivity. Results:①Got a dominant epitope at amino acid 71-89,called A1 region; ②Western-bl ot assay suggested that mSAK, a mutant SAK without A1 region., didn't combine to the anti-SAK PcAbs.Conclusion:A dominant epitope of SAK was mapped successfully with a simple,effective method . [
